Compositions and methods for treating coccidiosis



United States Patent 3,155,572 COMPOSITIONS AND METHODS FOR TREATINGCOCCIDIOSIS Edward F. Rogers, Middletown, and Lewis H. Sarett,

Princeton, N.J., assignors to Merck & Co., Inc., Rahway, NJ., acorporation of New Jersey No Drawing. Filed Apr. 13, 1959, Ser. No.805,738 9 Claims. (Cl. 167-53.1)

This invention relates generally to methods for the treatment ofcoccidiosis. More specifically, it is concerned with chemical compoundswhich are useful in the prevention and the cure of that disease. Stillmore specifically, it is concerned with quinolinium and isoquinoliumquaternary salts which are effective in controlling coccidiosis when fedin small amounts to poultry. It is further concerned with novelcompositions containing such substances.

Coccidiosis is a common and widespread poultry disease caused by severalspecies of protozoan parasites of the genus Eimeria, such as E. tenella,E. necatrix, E. acervulina, E. maxima, E. hagani and E. brunetti. E.tenella is the causative agent of a severe and often fatal infection ofthe ceca of chickens which is manifested by ex tensive hemorrhage,accumulation of blood in the ceca, and the passage of blood in thedroppings. E. necatrix as well as other species attack the smallintestine of the chick causing what is known as intestinal coccidiosis.Related species of coccidia such as E. melagridis and E. adenoides arecausative organisms of coccidiosis in turkeys. When left untreated, thesevere forms of coccidiosis lead to poor weight gain, reduced feedefficiency and high mortality in fowl. The elimination or control ofcoccidiosis is, therefore, of paramount importance to the poultryraising industry.

It has now been found that certain quinolinium and isoquinoliniumcompounds are highly active against the protozoa responsible forcoccidiosis. One object of the invention is to provide such compounds.Another object is to provide novel compositions containing suchsubstances as an active ingredient. A further object is the provision ofanimal feeds and feed supplements containing these quaternary salts.Other objects will be apparent from the following discussion:

According to this invention it has been found that1-(2-loweralkyl-4-amino-5-pyrimidylmethyl) quinolinium andisoquinolinium quaternary salts, and particularly those compoundswherein the quinolinium ring is substituted with 1 or 2 lower alkylgroups, are very effective in preventing and treating coccidiosis. Thepreferred compounds of our invention are quaternary salts of theformulae- CH N N a 0X- R N (R) n wherein R and R are lower alkylradicals, n has a value of 1 or 2, X is an anion, and b and c arepositive numbers having values such that the positive charge of b molesof and cation is neutralized by c moles of anion X. Thus when X is amonovalent anion such as a halide, b is 1 and c is 2. When more than onelower alkyl group is present in .the quinoline ring, i.e., when n is 2or3, such lower alkyl groups may be. the same or different.

As described more fully below, these anticoccidial compounds areprepared -by reaction of a 2-1ower-alkyl-4- amino-S-substitutedmethylpyrimidine with an appropriately alkylated quinoline or isoquinoline. Itwill be apparent from the above structural formulae that theanticoccidial compounds described herein may be considered assubstituted quinolines or isoquinolines with such heterocyclic nucleussubstituted at the 1-position by a2-loweralkyl-4-amino-S-pyrimidylmethyl radical; in the preferredcompounds, it is further substituted with one or two lower alkyl groups,such as methyl, ethyl, propyl, isopropyl, butyl and amyl groups,although the quaternaries wherein the quinoline or isoquinoline ring isunsubstituted are within the purview of this invention. As examples ofquinoline and isoquinoline compounds that may be quaternized with a2-lower-alkyl-4-amino-5-substitutedmethyl pyrimidine to give highly.active coccidiostat compounds there may be mentioned Z-methyl quinoline,4- methyl quinoline, 6-methyl quinoline, 6-propyl quinoline, 3-methy1isoquinoline, 1,4-dimethyl isoquinoline, quinoline, isoquinoline and thelike.

The loweralkyl group present in the 2-position of the4-amino-5-substimtedmethyl pyrimidine may be one such as a methyl,ethyl, propyl, isopropyl, butyl or amyl group. The lower alkyl groupspresent in these quaternary salts need not, of course, be the same inany particular compound.

The quaternary anion of our anticoccidial substances may be an inorganicanion such as chloride, bromide, iodide, nitrate, sulfate, phosphate andthe like, or the anion of an organic acid such as citric, tartaric,acetic, picric, stearic, succinic, benzoic, phthalic, phenoxyacetic,embonic, abietic, Z-naphthalene sulfonic or ethylenediamine tetraceticacids. It may also be the anion of a polymer such as a polyphosphate ofpolystyrenesulfonate ion. The nature of the anion is not critical andany anion may be employed as long as it is not unduly toxic for thepoultry. It will be realized that an acid addition salt of the primaryamine group present in these compounds will also be formed concurrentlywith the quaternary salt.

These substances are conveniently fed to poultry as a component of thefeed of the animals although they may also be given dissolved orsuspended in the drinking water. According to one aspect of theinvention, novel compositions are provided in which a quinolinium orisoquinolinium quaternary salt is present as an active anticoccidialingredient. Such compositions comprise the quaternary salts intimatelydispersed in or admixed with an inert carrier or diluent. By an inertcarrier is meant one that is nonreactive with respect to the quaternaryand that may be administered with safety to the animals. The carrier ordiluent is preferably one that is or may be an ingredient of the animalfeed.

The compositions which are a preferred feature of the invention are theso-called feed supplements in which the active ingredient is present inrelatively large amounts and which are suitably for addition to thepoultry feed directly or after an intermediate dilution or blendingstep. Examples of carriers or diluents suitable for such compositionsare solid orally ingestable carriers such as distillers dried grains,corn meal, citrus meal, fermentation residues, ground oyster shells,Attapulgus clay, wheat shorts, molasses solubles, corn cob meal, ediblevegetable substances, toasted dehulled soya flour, soybean mill feed,antibiotic mycelia, soya grits, crushed limestone and the like. Thequaternary salts are intimately dispersed or a admixed throughout thesolid inert carrier by methods such as grinding, stirring, milling ortumbling. By selecting proper diluents and by altering the ratio ofcarrier to active ingredient, compositions of any desired concentrationmay be prepared. Formulations containing from about 1% to about 40% byweight, and preferably from about 225% by weight of active ingredientare particularly suitable for addition to poultry feedstuffs, andcompositions containing from about 5-15% by weight of coccidiostat arevery satisfactory. The active compound is normally dispersed or mixeduniformly in the diluent but in some instances may be sorbed on thecarrier. The optimal concentration of coccidiostat in these feedsupplements will depend to some extend on the particular compoundemployed. Since it is convenient for the feed manufacturer to use aboutone pound of feed supplement for each ton of finished feed, the

preferred concentration of any one of our coccidiostats in a feedsupplement is partly a function of the level of active ingredientdesired in the finished feed.

Typical feed supplements containing quaternary salts representative ofour invention dispersed in an inert carrier include Lbs.

A. 1-(2-ethyl-4-amino-5-pyrimidylmethyl)-4-methy1 quinolinium chloridehydrochloride 6.0 Wheat standard middlings 94.0

B. l-(2 methyl 4 amino-S-pyrimidylmethyl)-isoquinolinium bromidehydrobromide 10.0 Corn distillers dried grains 90.0

. 1 (2 methyl 4 amino-S-pyrimidylmethyl)-4- methyl quinolinium bromidehydrobromide 15.0 Wheat standard middlings 85.0

F. 1 (2 methyl 4 amino-S-pyrimidylmethyl)-6- methyl quinolinium chloridehydrochloride 40.0 Soya grits 60.0

G. l-(2-ethyl-4-amino-S-pyrimidylmethyl)-2-methyl quinolinium sulfateFermentation residues 50.0 Wheat shorts 45.0

These and similar feed supplements are prepared by uniformly mixing thequaternary salt with the carrier or carriers.

The feed supplements of the type illustrated hereinabove are usuallyfurther diluted with materials such as corn meal or soybean meal beforebeing incorporated in the animal feed. In this intermediate processingstep the level of coccidiostat in the carrier is brought down to fromabout 0.1% to about 1.0% by weight. This dilution serves to facilitateuniform distribution of the substance in the finished feed. The finishedfeed is one that contains a source of fat, protein, carbohydrate,minerals, vitamins and other nutritional factors.

The amount of quinolinium or isoquinolinium quaternary salt requiredfoi' control of coccidiosis in poultry will, of course, vary somewhatwith the specific compound employed. The compounds of the invention areeffective in preventing the disease when administered at levels of lessthan about 0.05% by weight of the feed. Generally, good prophylacticresults are obtained when the quaternary is fed in amounts of from about0.0005% to about 0.05% by weight of the total feed consumed; for bestresults it is preferred that the poultry feed contain between about0.0025% and 0.025% by weight of quaternary salt. When the quinolinium orisoquinolinium salts are to be employed as therapeutic agents, thehigher levels are used for relatively short periods of time. Thus,

4 concentration of about 0.02% to 0.05% by weight of the feed may beadvantageously administered in treating an established outbreak ofcoccidiosis. It will be realized that the optimum dose level will varyslightly with the particular compound being employed.

Many of these quinolinium quaternary salts are advantageouslyadministered by way of the drinking water of the birds. This method oftreatment is often employed in the therapeutic use of our compoundssince poultry infected with coccidiosis are apt to consume less solidfeed than normal birds. The water-soluble quaternary salts may be addeddirectly to the drinking water. Alternatively, water-soluble powders maybe prepared, in which the coccidiostat is intimately admixed with asuitable carrier, such as dextrose or sucrose, and these powders addedto the drinking water of poultry as necessary. Such water-solublepowders may contain any desired concentration of coccidiostat, andprepartions containing from l-25% by weight of quaternary compound aresuitable in reacting our invention.

The compounds described herein are synthesized by reacting anappropriately substituted pyrimidine and a quinoline or isoquinolinecompound. According to one process, a 2-loweralkyl-4-amino-5-halomethylpyrimidine dihydrohalide, in which the halogen is bromine 'or chlorine,is reacted directly with the quinoline or isoquinoline. We prefer toemploy organic solvents inert under the reaction conditions such aslower alkanols, acetonitrile or an N,N-diloweralkyl alkanamide as thereaction medium. The temperature is not critical and it is convenient tocarry out the process at about room temperature. After a short time thequaternary salt crystallizes and is recovered by known techniques suchas filtration or centrifugation.

Although the S-halomethyl pyrimidines are generally most convenientlyemployed for the quaternization, we may also employ an ester of2-loweralkyl-4-amino-5- hydroxymethyl pyrimidine. Suitable esters arethe methylsulfite, p-toluenesulfonate and nitrate esters. Thequatemization may be conducted so that the particular salt desired fortreating coccidiosis is obtained directly. Alternatively, the quaternarysalt recovered from the synthetic reaction medium may be convenientlymetathesized to another salt by techniques known in the art.

The following examples are given for purposes of illustration and not byway of limitation:

Example 1 (a) Forty ml. of 4-methyl quinoline was added to a suspensionof 20 grams of 2-ethyl-4-amino-5-bromomethylpyrimidine dihydrobromide inml. of acetoni- Example 2 (a) To a suspension of 30 grams of2-methyl-4-amino- S-bromomethylpyrimidine dihydrobromide in ml. ofacetonitrile was added at room temperature 25 grams of3-tnethylisoquinoline. In a short time l-(2-methyl-4-ammo-S-pyrimidylmethyl)-3-methyl isoquinolinium bromide hydrobromidebegan to crystallize. The mixture was allowed to stand at roomtemperature for about 15 hours. The quaternary salt was filtered off andrecrystallized from aqueous acetone. It had a melting point of 247 C.(dec.).

Example 3 (a) The procedure of Example 1(a) was repeated usingas thereactants 15 grams of 2-ethyl-4-amino-5- bromomethylpyrimidinedihydrobromide and 25 ml. of 6-methyl quinoline. There was produced1-(2-ethyl-4- amino-6-pyrimidylmethyl)-6-methylquinolinium bromidehydrobromide. After recrystallization from aqueous acetone thequaternary salt had a melting point of 283 ,C. (dec.).

(b) When the method of Example 1(a) was carried out using '15 grams of2-ethyl-4-amino-5-bromoethyl- .pyrimidine dihydrobromide and 20 ml. of7-methyl quinol1ne as the reagents, 1- (2 ethyl 4amino-5-pyrim1dylmethyl) -7-methyl quinolinium bromide hydrobromide wasproduced. The product melted at 229 C. (dec.) after recrystallizationfrom aqueous acetone.

When Example 3(b) was carried out using 2- methyl quinoline in place of7-methyl quinoline, 1-(2- ethyl-4-amino-S-pyrimidylmethyl)-2-methylquinolinium bromide hydrobromide, melting point 195 C. (dec.) wasproduced.

Example 4 (a) To 30 grams of 2-methyl-4-amino-5-bromomethyl pyrimidinedihydrobromide in 80 ml. of methanol was 'added'90 ml. of 4-methylquinoline. The mixture was allowed to stand at about room temperaturefor about '15 hours. At the end of this time the crystalline 1-(2-methyl-4-amino-5-pyrimidylmethyl)-4-methyl quinolinium bromidehydrobromide was recovered by filtration and recrystallized from aqueousethanol. It had a melting point of 257 C. (dec.).

(b) Five gramsof the above material was dissolved in 20 ml. ofconcentrated hydrochloric acid, and the resulting solution dilutedcarefully with 400 ml. of acetone. The crystalline material thusobtained was redissolved in 20 ml. of concentrated hydrochloric acid andthe solution diluted with 400 ml. of fresh acetone. The resultingcrystalline 1-(2-methyl-4-aminO-S-pyrimidylmethyl)-4-methyl quinoliniumchloride hydrochloride was filtered and dried. It melted at 253 C.(dec.).

Example 5 (a1) 75 ml. of Z-methyl quinoline and 25 grams of 2-methyl-4-amino-5 bromomethylpyrimidine dihydrobromide were reactedtogether in 75 ml. of methanol. 1-('2- methyl-4-amino-5-pyrimidylmethyl)-2-methyl quinolinium bromide hydrobromide crystallized. 0nrecrystallization from aqueous ethanol and drying, it melted at 243 C.(dec.).

(b) Reaction of 20 grams of 2-methyl-4-amino-5-bromomethylpyrimidinedihydrobromide and 20 ml. of 6- methyl quinoline under the conditions ofExample 5 (w) gave 1-(Z-methyl-4-amino-5-pyrimidylmethyl)-6 methylquinolinium bromide hydrobromide, melting point 293 C. (dec.).

(c) When the procedure of Example 5 (a) was repeated in 100 ml. ofmethanol using 50 m1. of 7-methyl quinoline in place of 2-methylquinoline, there was produced l-(2-methyl-4-amino-5-pyrimidylmethyl)-7-methyl quinolinium bromidehydrobromide, melting point 241 C. (dec.).

' Example 6 (a) To a solution of 30 grams of 2*methyl-4-amino-5-bromomethylpyrimidine dihydrobromide in 500 ml. of ethanol at roomtemperature was added 70 ml. of isoquinoline. The mixture was allowed tostand overnight at about room temperature. The resulting crystalline 1-(2-methyl-4-amino-5 pyrimidylmethyl) isoquinolinium bromide hydrobromidewas recovered by filtration and dried. It melted at 256 C. (dec.).

(b) 160 ml. of quinoline was added to a solution of grams ofZ-methyl-4-amino-5-bromomethylpyrimidine dihydrobromide in 500 ml. ofethanol. After a short time crystals of1-(2-methyl-4-aminoJ-pyrimidylmethyl) quinolinium bromide hydrobromideappeared. These were recovered by filtration and treated as in Example4(b) to form the chloride hydrochloride, melting point 271 C. (dec.).

Example 7 The anticoccidial activity of the compounds of this inventionis determined by the following method:

Straight run White Leghorn chicks, in groups of three each, were weighedand placed in cages with wire floors. They were fed ad libitum astandard laboratory basal ration in which graded levels of thequaternary salt were intimately dispersed or blended. Normal andinfected control birds were fed basal ration containing no quaternarysalt. One the second day of test, the chicks were each orally inoculatedwith 50,000 sporulated oocysts of Eimerl'a tenlla. Papers under thecages were examined on the sixth, seventh and eighth days of assay forthe presence or absence of bloody droppings. A score of 0 was given ifno bloody droppings were observed; scores of 1, 2 or 4 were assigned for1-3, 4-6 and 6 bloody spots, respectively. 'On the eighth day of assay,the surviving birds were weighed, sacrificed and examined grossly forcecal coccidiosis lesions. Normal ceca were scored 0, and ceca withdetectable, moderate or maximal lesions were scored 1, 2 and 4,respectively. When a bird died and cecal coccidiosis lesions werepresent, a score of 5 was recorded. If the total of the two scores was0-5, the compound under test was rated active; if the total score was 6or more the compound was rated inactive at the concentration tested.

The following compounds were active at the dose level indicated.Compound: Percent in feed l-(2-methyl-4 amino 5 pyrimidylmethyl)quinolinium bromide hydrobromide 1-(2-methyl-4-amino-5-pyrimidylmethyl)6- methyl quinolinium bromide hydrobromide1-(2-methyl-4-amino-S-pyrimidylmethyl) 7- methyl quinolinium bromidehydrobromide 1-(2-methyl-4-amino-5-pyrimidylmethyl) 2- methylquinolinium bromide hydrobromide 1-(2-methyl-4-amino-5-pyrimidylmethyl)4- methyl quinolinium bromide hydrobromide 0.0125 l-(2-ethyl-4-amino-5pyrimidylmethyl) 6- methyl quinolinium bromide hydrobromide 0.0125l-(2-ethyl-4-amino-S pyrimidylmethyl) 7- lnethyl quinolinium bromidehydrobromide 0.006 l-(2-ethyl-4-amiuo-5 pyrimidylmethyl) 4- methylquinolinium bromide hydrobromide 0.002l-(Z-ethyl-4-amino-5-pyrimidylmethyl) isoquinolinium bromidehydrobromide 0.025 I-(Z-ethyM-amino-S pyrimidylmethyl) 3- methylisoquinolinium bromide hydrobromide a 0.006.1-(2-methyl-4-amino-S-pyrimidylmethyl) 3- methyl isoquinolinium bromidehydrobromide 0.002 I(2-1nethyl-4-amino-5-pyrimidylmethyl) isoquinoliniumbromide hydrobromide 0.025

Example 8 The 2-loweralkyl-4-amino-5 halomethyl pyrimidines employed inmaking the quaternary compounds of this invention are prepared in thefollowing manner:

(a) 2-I0weralkyl-4-amino-5-cyanopyrimidine.-A slurry of 54.7 grams ofbutyramidine hydrochloride and 55 ml. of absolute ethanol is treated atabout 5 C. with a solution of 11 grams of sodium in 220 ml. of absoluteethanol. The resulting solution is added with stirring at l-12 C. over a30 minute period to 53.7 grams of ethoxymethylenemalononitrile in 54 ml.of absolute ethanol. The resulting mixture is stirred at 0 C. for sixhours and then at room temperature for about 12 hours. The mixture isthen filtered, evaporated to dryness in vacuo and the residue treatedwith water. The alcoholic and aqueous solution precipitates arecombined, washed with water and dried in air. The product isrecrystallized from alcohol to give2-n-propyl-4-amino-S-cyanopyrimidine, melting point 158-160 C.

When the above reaction is carried out with isobutyrnmidine there isobtained 2-isopropyl-4arnino-5-cyanopyrimidine, melting point ISO-151 C.When amylamidine is employed as starting material the end product is2-butyl-4-amino-S-cyanopyrimidine, melting point 143- 147 C. Whenhexylamidine is utilized as starting compound, there is obtained2-amyl-4-amino-S-cyanopyrirnidine, melting point 149-150 C.

(b) 2-IoweraIkyl-4-amino aminomethylpyrimidine dihydrochloride.l6.2grams of 2-n-propyl-4-amino-5- cyanopyrirnidine is reduced at about 40lbs. pressure in 200 ml. of methanol in the presence of 26 grams ofammonia and one tablespoon of Raney nickel. The drop in pressure isabout 36.5 lbs. The reaction mixture on completion of the reduction isconcentrated to a syrup after filtering oil the catalyst. The residuethus obtained is acidified with dilute hydrchloric acid and concentratedto a crystalline mass. 0n recrystallization from methanolacetone thereis obtained 2-n-propyl-4-amino-5-aminomethylpyrimidine dihydrochloride,melting point 220- 222 C.

When the 2-isopropyl, 2-butyl and 2-amyl-4-amino-5- cyanopyrirnidineobtained as described above are used as starting materials in thisreduction, there are obtained respectively 2-isopropyl-4-amino 5aminomethylpyrimidine dihydrochloride, melting point 257-260 C.,2-butyl- 4-arnino-5-aminomethylpyrimidine dihydrochloride, melting point221-223 C., and Z-amyl-4-amino-5-aminomethylpyrimidine dihydrochloride,melting point 188-489 C.

(c) 2-l0weralkyl-4-amino 5 hydroxy methyl pyrimidirr.-TW1V grams of2-n-propyl-4-amino-5-aminomethylpyrimidine dihydrochloride in 50 ml. ofwater is treated at 50-60 C. with a solution of 3.5 grams of sodiumnitrite in 30 ml. of water. The sodium nitrite is added dropwise over a45-minute period. The heating is continued for an additional two hours,and the reaction mixture then made alkaline with sodium carbonate andextracted with butanol. The butanol extract is evaporated to dryness andthe residue crystallized from acetone to give 2-n-propyl 4 aminoS-hydroxymethylpyrimidine, melting point 115-116 C.

When the Z-isopropyl, 2-butyl and 2-amyl-4-amino-S-aminomethylpyrimidine dihydrochlorides obtained as in Part b above isutilized in this reaction in place of the Z-n-propyl compound, there areobtained 2-isopropyl-4- amino 5 hydroxymethylpyrimidine,2-butyl-4-amino-5- hydroxymethylpyrimidine and2-amyl-4-amino-5-hydroxymethylpyrimidine.

(d) 2 laweralkyl 4 amino-S-bromo methyl pyrimidine-The2-n-propyl-4-amino 5 hydroxymethylpyrimidine obtained in Part 0 above isdissolved in 15 ml. of acetic acid saturated with hydrogen bromide, andthe mixture allowed to stand at room temperature for about 15 hours.Z-n-propyl-4-amino-5-bromomethylpyrimidine dihydrobromide crystallizesand is recovered by filtration and washed with ether. The material issubstantially pure and may be used directly in preparing the quaternarysalts of this invention.

The other 2-loweralkyl-4-amino-5-hydroxymethylpyrimidines describedabove are treated in like manner with hydrogen bromide to give2-isopropyl-4-amino-5-bromomethylpyrimidine dihydrobromide, meltingpoint 191- 192 C., 2-butyl-4-amino-S-bromomethylpyrimidinedihydrobromide, melting point -150 C., and 2-amyl-4-amino-S-bromomethylpyrimidine dihydrobromide.

Any departure from the above description which conforms to the presentinvention is intended to be included within the scope of the claims.

What is claimed is:

1. A composition useful against coccidiosis that comprises a solidnutritive poultry feed additive having intimately dispersed therein aquaternary salt selected from the class consisting of a compound havingthe formulaand compounds having the formulawherein R and R are loweralkyl groups, X is a nontoxic anion, and b and c are positive numbers ofsuch value that b moles of cation are neutralized by c moles of anion X.

2. A poultry feed supplement composition that com prises a solidnutritive poultry feed additive having dispersed therein from about 1%to about 40% by weight of an acid addition salt of a1-(2-lower-alkyl-4-amino-5- pyrimidylmethyi)-quinolinium quaternarysalt, wherein the anion of the salt is a non-toxic anion.

3. A poultry feed supplement composition that comprises a solidnutritive poultry feed additive having dispersed therein from about 1%to about 40% by weight of an acid addition salt of s1-(2-lower-alky1-4-smino- 5 pyrimidylmethyl) isoquinolinium quaternarysalt, wherein the anion of the salt is a non-toxic anion.

4. A composition useful against coccidiosis that comprises a solidnutritive poultry feed additive having dispersed therein from about 1%to about 40% by weight of l-(2-ethyl-4-amino S pyrimidylmethyl) 4 methylquinolinium halide hydrohalide.

5. A composition useful against coccidiosis that comprises a solidnutritive poultry teed additive having dispersed therein from about 1%to about 40% by weight of1-(2-methyl-4-smino-5-pyrimidylmethyl)-3-methyl isoquinolinium halidehydrohslide.

6. A poultry feed supplement composition that comprises a solidnutritive poultry feed additive having dispersed therein from about 1%to about 40% by weight of an acid salt of al-(2-lowerslltyl-4-smino-5-pyrimidylmethyl)-quinolinium quaternary salt,wherein the quincline ring is mono-lower slkylsted and the anion at thesalt is a non-toxic anion.

7. A poultry feed supplement composition that comprises a solidnutritive poultry feed additive having dispersed therein from about 1%to about 40% by weight of an acid salt of sl-(2-lowerslkyl-4-amino-5-pyrimidylmethyl)-isoquinolinium quaternarysalt, wherein the isoquinoline ring is mono-lower slkylsted and theanion of the salt is a non-toxic anion.

8. An animal teedstuft having dispersed therein from about 0.0005 toabout 0.05% by weight of a quaternary salt selected from the classconsisting of a compound having the formula t N CH :;\I\/ cX- R, k (runand compounds having the formulaon i: N

cX- MLN (m b wherein R and R are lower alkyl groups, X is a nontoxicanion, n has a value of 0 to 2 inclusive, and b and c 20 1y dispersedtherein from about 0.0005% to about 0.05%

by weight of a quaternary salt selected from the class consisting ofcompounds having the formulaand compounds having the formulawherein Rand R are lower alkyl groups, X is a nontoxic anion, n has a value of *0to 2 inclusive, and b and c are positive numbers of such value that bmoles of cation are neutralized by c moles of anion X.

References Cited in the file of this patent Taizo: Chem. Abstracts, vol.50, 1956, p. 1010G. Chem. Abst. Subject Index, vol. 51, 1957, p. 20708.

Patent No. 3,155,572 November 3, 1964 Edward F. Rogers et a1.

tified that error appears in the above numbered pat- Patent should readas It is hereby cer ion and that the said Letters ent requiring correctcorrected below.

Column 9, lines 3 to 9. the formula should appear as shown below insteadof as in the patent:

Signed and sealed this 16th day of March 1965,

(SEAL) Attest:

- EDWARD J. BRENNER ERNEST we SWIDER A ttesting Officer Commissioner ofPatents

1. A COMPOSITION USEFUL AGAINST COCCIDIOSIS THAT COMPRISES A SOLIDNUTRITIVE POULTRY FEED ADDITIVE HAVING INTIMATELY DISPERSED THEREIN AQUATERNARY SALT SELECTED FROM THE CLASS CONSISTING OF A COMPOND HAVINGTHE FORMULA-